CHAPTER 1 (FLUOROMETRY)

Question 1

fluorometry utilises ___ which specifically ___

Answer 1

1. fluoroscent dyes
2. bind DNA or RNA

Question 2

what does it require?

Answer 2

1. negative control
2. standard of known concentration

Question 4

whats the function of negative control

Answer 4

1. verify that theres no fluorescence bg
2. ensure specificity of measurement
3. to set the zero point on the fluorometer

Question 5

how does the fluorometry works?

Answer 5

1. The fluorometer shines light on the sample (excitation) and then measures level of fluorescent light being emitted to the
   side (at a 90º angle) of the excitation light beam.
2. The fluorescent dyes are relatively specific to nucleic acids as opposed to protein and other cellular components.
3. The fluorescence of the dyes increases when they bind nucleic acids.

Question 6

fluorometry is ____ than spectrophotometric absorbance

less susceptible to_____

Answer 6

1. 1000x more sensitive
2. protein and RNA contamination

Question 7

it does not give a crude measurement of____ nor does it assure that _____

Answer 7

1. purity
2. the DNA or RNA is not degraded (like suze determination by gel electrophoresis)

Question 8

Do not use glass (spectrophotmetry) cuvettes in a fluorometer. why?

Answer 8

the frosted glass on the side of the cuvette interferes with detection of fluorescent light.